Isolation And Purification Of Xyloglucanase From Snail Livers

Xyloglucanase is the enzyme that can hydrolate xyloglucan. It is widely found in bacteria, fungi and mollusks of nature. Not only does it have a good prospect in fodder, food and pharmaceutical industry, but also it has been focused on by domestic and foreign counterparts and have gotten some important achievements.This paper aimed to study xyloglucanase of snail liver, its extraction, isolation and purification, identification and some physical and chemical properties have been researched, which will provide technical support to the sequencing of xyloglucanase, gene cloning and the monoclonal of antibodies.After washing, weighing, crushing snail stomach, prime Mang and liver tissue, we constructed the SDS-PAGE electrophoresis of these tissues by the method that homogenate snail tissue with phosphate extraction buffer(concentration of 1/15M,pH is 5.29, the ratio of tissue to buffer is 1:4) in 4℃, then do the ammonium sulfate fractionation procedure in 0℃and SDS-PAGE.The xyloglucanase of different parts of the liver were extracted with different ratios of Na2HPO4-KH2PO4 extraction buffer, followed by ammonium sulfate fractionation, electrophoretic desalinization and the determination of Enzyme activity, we draw the results as follows: Each part of snails'livers has the Enzyme activity generally, but there exsit differences in various parts, the third part is the highest, then the first part, the second part and the fourth part are the lowest; Ammonium Sulfate with thirty percent saturation are able to gain the highest specific activity.Xyloglucanase was extracted with Na2HPO4-KH2PO4 extraction buffer, followed by ammonium sulfate fractionation, Sephadex G-150 gel filtration and DEAE- Sephadex A-50 exchange resin purification(specific activity reached 28.43U/mg, purified multiple was 7.60 times, the recovery rate is 6.37%), polyacrylamide gel electrophoresis,we gain the pure xyloglucanase of snail liver. According to some physical and chemical characteristics, the enzyme's relative molecular weight is 22.7KDa, and the optimum temperature and pH is 55℃and 5.6 respectively.The program and the technology routes of the study for isolation and purification can satisfy the need for extraction, isolation and purification of snail xyloglucanase. Snail xyloglucanase gained by electrophoresis also can meet the requirements of sequencing and monoclonal antibody.